Journal: Journal of Medicinal Chemistry

Article Title: Nanoscale Direct-to-Biology Optimization and Structural Insights into Selective S. aureus TrmD Inhibitors

doi: 10.1021/acs.jmedchem.5c02323

Figure Lengend Snippet: Definition of starting points for the development of S. aureus TrmD inhibitors. (A) Crystallographic overlay of literature-known inhibitors 1 (PDB: 4MCD ) and 2 (PDB: 4MCC ) in the H. influenzae SAM-binding pocket. Ligands and amino acids are shown in stick representation; polar interactions are shown as yellow lines, and water as red spheres. The carbon atoms of 1 are shown in pink and those of compound 2 in light green. Interacting residues are displayed in dark green. (B, C) Structures and affinities of inhibitors 1 (B) and 2 (C) binding to S. aureus TrmD determined by ITC, incl. thermograms, stoichiometry, and signature plots (data for H. influenzae TrmD in Figure S2 ). Due to low enthalpy signals, a low c -titration was performed for inhibitor 1 , resulting in N < 1.

Article Snippet: The enzymatic reactions were started by the addition of SAM as a mixture of 3 H-SAM (Hartmann Analytics, S. aureus TrmD: 0.5 μM, Trm5: 2 μM) and cold SAM (New England Biolabs, S. aureus TrmD: 0.5 μM, Trm5: 98 μM) to a final activity of 0.04 μCi μL –1 (TrmD) or 0.16 μCi μL –1 (Trm5).

Techniques: Binding Assay, Titration